Hello colleagues!
I performed RNA extraction for my cells using TRIzol, and when I measured the yield using nanodrop, I got two peaks of RNA.
I read that it is because of phenol contamination. These samples are extremely important to me. Can you please suggest any way to eliminate the contamination?
And are there any tips to avoid this in future?
Thanks.
this is because of contamination from other cellular materials which absorbs lights at same wavelength. I am not sure but you can try running it on gel and if 28S rRNA band appears, as it should be in case of eukaryotes, you can try RNA gel extraction and purification with standard kits available in the market.
Yes, it is a common UV absorptance at 220 nm due to TRIzol phenol reagents contamination. Make sure you did a properly cleaned up.
You may want to try spin columns from some RNA purification Mini Kit.
They have provided options for TRIzol or TRIzol-free clean-up protocol.
Good luck!
230 nm is for organic solvents. Unfortunately you are not removing wheither ethanol or phenol completely.
You can add one extra wash step, if you still have the peak; then you need to increase drying time of ethanol to provide sufficient evaporation of ethanol.
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